Improvement of Na+-K+ ATPase Activity and Gene Expression Associated with Glomerular Ultrastructural Protection in Diabetic Nephropathy by Acacia senegal: Role of Oxidative Stress Disturbances / Mejora de la actividad Na+-K+ ATPasa y la Expresión Génica Asociada con la Protección Ultraestructural Glomerular en la Nefropatía Diabética por Acacia senegal: Papel de las Alteraciones del Estrés Oxidativo

SUMMARY: This study assessed the effects of Acacia Senegal (AS) combined with insulin on Na+/K+-ATPase (NKA) activity and mRNA expression, serum glucose, renal function, and oxidative stress in a rat model of diabetic nephropathy (DN). Sixty rats were equally divided into six groups: normal control, normal+AS, diabetic (DM), DM+insulin, DM+AS, and DM+insulin+AS groups. Diabetes mellitus (type 1) was induced by a single injection of streptozotocin (65 mg/kg), and insulin and AS treatments were carried until rats were culled at the end of week 12. Serum glucose and creatinine levels, hemoglobin A1c (HbA1c) were measured. Renal homogenate levels of NKA activity and gene expression, malondialdehyde, superoxide dismutase (SOD), catalase and reduced glutathione (GSH) were evaluated as well as kidney tissue histology and ultrastructure. Diabetes caused glomerular damage and modulation of blood and tissue levels of creatinine, glucose, HbA1c, malondialdehyde, NKA activity and gene expression, SOD, catalase and GSH, which were significantly (p<0.05) treated with AS, insulin, and insulin plus AS. However, AS+insulin treatments were more effective. In conclusion, combined administration of AS with insulin to rats with DN decreased NKA activity and gene expression as well as oxidative stress, and improved glycemic state and renal structure and function.

Este estudio evaluó los efectos de Acacia senegal (AS) combinada con insulina sobre la actividad Na+/K+- ATPasa (NKA) y la expresión de ARNm, la glucosa sérica, la función renal y el estrés oxidativo en un modelo de nefropatía diabética (ND) en ratas. Sesenta ratas se dividieron equitativamente en seis grupos: control normal, normal+AS, diabética (DM), DM+insulina, DM+AS y DM+insulina+AS. La diabetes mellitus (tipo 1) se indujo mediante una única inyección de estreptozotocina (65 mg/kg), y los tratamientos con insulina y AS se llevaron a cabo hasta que las ratas fueron sacrificadas al final de la semana 12. Se midieron niveles séricos de glucosa y creatinina, hemoglobina A1c (HbA1c). Se evaluaron los niveles de homogeneizado renal de actividad NKA y expresión génica, malondialdehído, superóxido dismutasa (SOD), catalasa y glutatión reducido (GSH), así como la histología y ultraestructura del tejido renal. La diabetes causó daño glomerular y modulación de los niveles sanguíneos y tisulares de creatinina, glucosa, HbA1c, malondialdehído, actividad y expresión génica de NKA, SOD, catalasa y GSH, los cuales fueron tratados significativamente (p<0,05) con AS, insulina e insulina más AS. Sin embargo, los tratamientos con AS+insulina fueron más efectivos. En conclusión, la administración combinada de AS con insulina a ratas con DN disminuyó la actividad de NKA y la expresión genética, así como el estrés oxidativo, y mejoró el estado glucémico y la estructura y función renal.

The effect of salinity on osmoregulation and development of the juvenile fat snook, Centropomus parallelus (POEY) / O efeito da salinidade sobre a osmoregulacao e o desenvolvimento do juvenil de robalo-peva, Centropomus parallelus (POEY)

Eurihaline fish support waters with different salt concentration. However, numerous studies have shown that salinity can affect fish development. Thus, the effect of salinity change from 20 to 5 and 35 on survival, weight, length, gill chloride cell ultrastructure and gill Na+, K+ ATPase activity was evaluated in Centropomus parallelus following short-term (6, 24 and 96 hours) and long-term exposure (30 and 60 days). Salinity did not affect C. parallelus survival, final weight and length. The quantity of chloride cells increased visibly at salinities of 5 and 35, with the cells exhibiting the typical features of uptake and secretory cells, respectively. Na+, K+ ATPase activity in the gill of the C. parallelus was significantly greater at a salinity of 5 than at a salinity of 20 or 35 after 96 hours, but not after 30 or 60 days. These results indicate that salinity change from high to low salt water induces gill chloride cell and Na+, K+ ATPase activity adaptations after short-term exposure. However, after long-term exposure at salinity 5, gill Na+, K+ ATPase activity is no more necessary at high levels. The increase in salinity to 35 does not induce significant change in gills. Juveniles of C. parallelus may thus be capable of acclimating to salinities of 5 to 35 for 60 days without significant effects on development.

Peixes eurihalinos suportam águas com diferentes concentrações de sal. Contudo, muitos estudos têm mostrado que a salinidade pode afetar o desenvolvimento do peixe. Portanto, o efeito da mudança de salinidade de 20 para 5 e 35 na taxa de sobrevivência, peso, comprimento, morfologia das células de cloreto branquiais e atividade da Na+, K+ ATPase foram avaliadas no Centropomus parallelus após curto (6, 24 e 96 horas) e longo tempo de exposição (30 e 60 dias). A salinidade não afetou a sobrevivência, o peso e comprimento final do robalo-peva. A quantidade de células de cloreto aumentou visivelmente nas salinidades 5 e 35, exibindo morfologias típicas de células que absorvem e secretam sal, respectivamente. A atividade da Na+, K+ ATPase nas brânquias do C. parallelus foi significativamente maior na salinidade 5 do que nas salinidades 20 ou 35 após 96 horas, mas não após 30 e 60 dias. Esses resultados indicam que a mudança de alta para baixa salinidade provoca adaptações nas células de cloreto e na atividade da Na+, K+ ATPase branquial em curto prazo. Contudo, após longa exposição na salinidade 5, a alta atividade da Na+, K+ ATPase branquial não é mais necessária. O aumento de salinidade para 35 não induz mudanças significativas nas brânquias. Portanto, juvenis de C. parallelus possuem a capacidade de aclimatação nas salinidades de 5 a 35 semefeitos significativos no desenvolvimento após 60 dias.

Hypertensive effects of the iv administration of picomoles of ouabain

Ouabain, an endogenous digitalis compound, has been detected in nanomolar concentrations in the plasma of several mammals and is associated with the development of hypertension. In addition, plasma ouabain is increased in several hypertension models, and the acute or chronic administration of ouabain increases blood pressure in rodents. These results suggest a possible association between ouabain and the genesis or development and maintenance of arterial hypertension. One explanation for this association is that ouabain binds to the α-subunit of the Na+ pump, inhibiting its activity. Inhibition of this pump increases intracellular Na+, which reduces the activity of the sarcolemmal Na+/Ca2+ exchanger and thereby reduces Ca2+ extrusion. Consequently, intracellular Ca2+ increases and is taken up by the sarcoplasmic reticulum, which, upon activation, releases more calcium and increases the vascular smooth muscle tone. In fact, acute treatment with ouabain enhances the vascular reactivity to vasopressor agents, increases the release of norepinephrine from the perivascular adrenergic nerve endings and promotes increases in the activity of endothelial angiotensin-converting enzyme and the local synthesis of angiotensin II in the tail vascular bed. Additionally, the hypertension induced by ouabain has been associated with central mechanisms that increase sympathetic tone, subsequent to the activation of the cerebral renin-angiotensin system. Thus, the association with peripheral mechanisms and central mechanisms, mainly involving the renin-angiotensin system, may contribute to the acute effects of ouabain-induced elevation of arterial blood pressure.

Effect of plant molluscicides on selected enzymes related to energy metabolism in biomphalaria arabica snails molluscan hosts to schistosoma mansoni in Saudi Arabia

Schistosomiasis is one of the most important human parasitic diseases. One of the possible methods for the control is through the molluscan intermediate host of the parasite. Biomphalaria arabica, molluscan hosts to Schistosoma mansoni in Saudi Arabia were treated with sublethal concentrations [LC25] of dry powdered leaves Solanum nigrum. Effect of plant on ectonucleotidases [NTPdases] [ADPase and ATPase], sodium/potassium adenosine triphosphatase [Na[+]/K[+] ATPase] and creatine kinase [CK] was traced. The plant molluscicide was potent in inhibiting the four investigated enzymes giving a percentage inhibition range between 45-55%. The effect of the inhibited enzymes on the compatibility of the snail hosts to schistosome parasite was discussed. In conclusion, the use of sublethal concentration of S. nigrum to disturb the biochemical profile of the snail hosts could be a promising and safe strategy to control the disease

Infusión continua de diurético en el manejo de la retención de líquido pulmonar del recién nacido / Continued diuretic infusion in the treatment of pulmonary liquid retention in newborns

Introducción. La retención del líquido pulmonar (RLP) es una patología benigna relativamente frecuente, que se presenta principalmente en niños de término, nacidos por cesárea y cuya fisiopatología consiste en retardo en la reabsorción del líquido pulmonar que ocurre en las primeras horas de vida y que puede dar manifestaciones clínicas variables. La furosemida administrada en bolos no ha tenido resultados alentadores en acelerar el proceso de reabsorción, en este estudio se aplicó el medicamento en infusión continua y se observó la evolución y duración de esta enfermedad. Material y métodos. Se estudiaron 40 recién nacidos de término con diagnóstico clínico y radiológico de RLP y se formaron dos grupos: el A con diurético en infusión continua y el B con manejo convencional. Resultados. No hubo diferencias significativas en cuanto a sexo, vía de nacimiento, calificación de Apgar, edad gestacional y peso al nacer. Se evaluaron electrolitos séricos pre y post.tratamiento en el grupo A, aunque estadísticamente no significativa. Fue más rápida la resolución en el grupo que se administró la infusión de furosemide (13.9 ñ 7.5 vs 23.6 ñ 10.3, P < 0.001). Conclusión. Aunque en este trabajo se comprobó resolución más rápida de la patología, ésta es de sólo 10 horas por lo que no se considera la aplicación de furosemide en forma rutinaria

Estructura molecular de los receptores de diuréticos luminales / Molecular structure of luminal diuretic receptors

El riñon juega un papel central en el mantenimiento del volumen y la composición del líquido corporal. En condiciones fisiológicas, la ingesta de sal y agua, día con día, es más o menos constante, por lo que la excreción urinaria de sodio es determinante en el mantenimiento del líquido corporal. esta función la lleva a cabo el riñon gracias a la integración de la ultrafiltración, en el glomérulo, y de la reabsorción tubular a lo largo de la nefrona. El control fino de la excreción urinaria de sodio se lleva a cabo gracias a la reabsorción tubular en la nefrona distal. En este sitio, con base en el requerimiento de iones (CI- y K+) y la sensibilidad a diuréticos, tres diferentes mecanismos de transporte de sodio se han identificado en la membrana apical: 1) los cotransportadores de Na+:K+:2CI- y Na+:CI- sensibles a derivados del ácido sulfamoílbenzoico (furosemide o bumetanida) en el asa ascendente de Henle; 2) el contrasportador de Na+:CI- sensible a la benzotiadiazina (o tiazidas) en el tíbulo distal; y 3) los canales de Na+ sensibles a amilorida en el túbulo colector. La inhibición de esta proteínas con diuréticos aumenta la excreción urinaria de Na+. Gracias a técnicas avanzadas en biología molecular, como la clonación por expresión funcional en ovocitos de Xenopus laevis, se ha identificado actualmente DNAc que codifica para miembros de las familias de estos receptores. En este trabajo se presenta una revisión sobre los avances en el conocimiento de la estructura primaria y de la relación entre la estructura y función de los receptores para diuréticos. Se discuten también las posibles consecuencias que estos hallazgos tendrán para el entendimiento en la fisiología y la fisiopatología del manejo renal del sodio

Studies on the effect of ethanol on the Na+, and the Na+,K+ATPase activities of plasma membranes of rat kidney proximal tubular cells

In the present work it was investigated the effect of 2 percent ethanol on the Na+ and on the Na+, K(+)-ATPase activities. The differential effect of the alcohol on the two ATPases (approximately 40 percent inhibition of the Na(+)-ATPase and approximately 10 percent inhibition of the Na+, K(+)-ATPase), is not due to a higher degree of denaturalization of the enzyme, nor to a faster effect of ethanol on the Na(+)-than on the Na+, K(+)-ATPase. Our results show that ethanol affects the selectivity of the Na+, K(+)-ATPase for Na+ and/or for K+, enhancing the Na+ affinity for the K+ sites, and/or reducing the K+ affinity for its own sites. This effect was not seen for the Na(+)-ATPase, indicating that 2 percent ethanol inhibits the two ATPases in a totally different way

Effects of Na+, K(+)-pump inhibitors on acetylcholine-induced relaxation in the rabbit aorta

The purpose of this study was to investigate the effects of inhibitors of the Na+, K(+)-pump and membrane depolarizing agents on endothelium-dependent acetylcholine-induced relaxation in the rabbit thoracic aorta. Aortic rings were prepared from the rabbit descending thoracic aorta and the contractility of the ring was measured in various conditions such as application of ouabain, exposure to K(+)-free Krebs-Henseleit solution and high K+. Ouabain or exposure to K(+)-free Krebs-Henseleit solution inhibited acetylcholine or sodium nitroprusside-induced relaxation. KCl also inhibited the acetylcholine or sodium nitroprusside-induced relaxation. These results suggest that the Na+, K(+)-pump may play a role in endothelium-dependent acetylcholine-induced relaxation.

Physiological, biochemical and molecular mechanisms of salt appetite control by mineralocorticoid action in brain

Adrenocortical hormone effects in the central nervous system depend on steroid interaction with intracellular receptors, which belong to a superfamily of ligand-activated transcription factors. Using a combination of biochemical and molecular biology techniques, we have demonstrated: 1. the localization of mineralocorticoid receptors in the brain, with highest density present in hippocampus, lateral septum and some amygdaloid nuclei; 2. the arousal of a mineralocorticoid-specific behavior such as salt appetite, coincident with inhibition of the biosynthesis/activity of (Na+K)ATPase in some amygdaloid and hypothalamic nuclei; 3. the modulation of the biosynthesis/activity of the sodium pump by glucocorticoids, although for these hormones changes are stimulatory, as shown in the spinal cord and brain; 4. the reported steroid effects on the (Na+K)ATPase constitute an important mechanism of control of nervous system function, involving behavior, changes in excitability and neurotropism